Quantification of X sperm by raman spectroscopy in percoll density gradient centrifuged buffalo semen
DOI:
https://doi.org/10.56825/bufbu.2023.4243847Keywords:
Bubalus bubalis, buffaloes, Raman spectroscopy, percoll, centrifugation, PBSAbstract
The present study was conducted to observe effect of percoll density gradient centrifugation of buffalo bull semen on quantity of X sperms. Ejaculates were collected by artificial vagina method. Semen with mass motility >+3 and progressive motility >70 % were selected for experiment. X sperm Enrichment of semen was done by discontinuous percoll density gradient centrifugation and three groups were formed ie Group 1 (3 layer 70%, 50% and 30%) Group 2 (7 layer 70%, 60%, 50%, 40%, 30%, 20% and 10%) Group 3 (7 layer 80%, 70%, 60%, 50%, 40%, 30% and 20%). Centrifugation of semen of three groups and control (fresh semen without gradients) was done. After centrifugation, the supernatant part was removed and the pellet of each group was used for X sperm enrichment assessment by Raman spectroscopy. Results revealed that X sperm enrichment was higher in the pellets of Group 2 followed Group 3, Group 1 and Control as Raman peaks on DNA specific bands corresponds to more number of x sperm were higher respectively.
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